Performance evaluation of nine reference centers and comparison of DNA extraction protocols for effective surveillance of Leishmania-infected Phlebotomine sand flies
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| Publication Date: | 2024 |
| Other Authors: | , , , , , , , , , , , , , , , , , , , , , , , , , , , |
| Format: | Article |
| Language: | eng |
| Source: | Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) |
| Download full: | http://hdl.handle.net/10362/178507 |
Summary: | Publisher Copyright: © 2024, Author. All rights reserved. |
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Performance evaluation of nine reference centers and comparison of DNA extraction protocols for effective surveillance of Leishmania-infected Phlebotomine sand fliesBasis for technical recommendationsPublic Health, Environmental and Occupational HealthInfectious DiseasesSDG 3 - Good Health and Well-beingPublisher Copyright: © 2024, Author. All rights reserved.Background Leishmaniasis, caused by Leishmania protozoan parasites transmitted by Phlebotomine sand flies, is a significant public health concern in the Mediterranean basin. Effective monitoring of Leishmania-infected sand flies requires standardized tools for comparing their distribution and infection prevalence. Consistent quantitative real-time PCR (qPCR) parameters and efficient DNA extraction protocols are crucial for reliable results over time and across regions. However, the absence of standardized technical recommendations for Leishmania DNA detection hinders effective surveillance. This study aimed to compare different DNA extraction protocols and conduct a qPCR-based External Quality Assessment (EQA) through a multicenter study involving nine reference laboratories, with a focus on optimizing Leishmania DNA detection in sand fly. Methodology/Principal findings EQA samples consisted of Leishmania infantum and L. major species, at concentrations ranging from 101 to 104 parasites/mL. All but one center detected all concentrations, demonstrating strong diagnostic proficiency. The ability to detect low concentrations highlighted the robustness of the qPCR assay used, though variations in Cq values indicated differences in sensitivity related to technical capabilities or DNA extraction kit performance. A comparative analysis of seven DNA extraction methods identified the EZ1 DSP Virus Kit and QIAamp DNA mini-kit as the most efficient, supporting their use in standardized protocols. The study also assessed the effects of lyophilization and shipment conditions, showing no significant compromise in Leishmania detection despite slight variations in Cq values. Experimentally infected sand flies were included to simulate field conditions, and all centers successfully detected positive samples with varying Cq values, probably reflecting differences in infection load. Conclusion and significance This study emphasizes the importance of standardized DNA extraction protocols and continuous quality assurance for accurate Leishmania DNA detection. The results highlight the superior performance of certain extraction kits and the need for ongoing technical training, essential for reliable leishmaniasis surveillance, particularly in field settings with low infection densities.Instituto de Higiene e Medicina Tropical (IHMT)Global Health and Tropical Medicine (GHTM)Laboratório Associado de Translacção e Inovação para a Saúde Global - LA Real (Pólo IHMT)Vector borne diseases and pathogens (VBD)RUNPrudhomme, JorianDelabarre, AymericAlten, BulentBerberoglu, UmutBerriatua, EduardoBongiorno, GioiaCristovão, José ManuelDavidovich-Cohen, MayaDi Muccio, TrentinaKasap, Ozge ErisozFiorentino, EleonoraKirstein, Oscar D.Kniha, EdwinMaia, CarlaMungan, MesutMuñoz-Hernández, ClaraNalcaci, MuhammedKaskan, Gizem OguzOzbel, YusufToz, Seray OzensoyParreira, RicardoPlatzgummer, KatharinaPolat, CeylanRisueño, JoséStudentsky, LioraVarol, GamzeWalochnik, JuliaYetişmiş, KardelenRobert-Gangneux, Florence2025-02-05T21:21:10Z2024-122024-12-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article15application/pdfhttp://hdl.handle.net/10362/178507eng1935-2727PURE: 108808671https://doi.org/10.1371/journal.pntd.0012543info:eu-repo/semantics/openAccessreponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiainstacron:RCAAP2025-02-10T01:39:36Zoai:run.unl.pt:10362/178507Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireinfo@rcaap.ptopendoar:https://opendoar.ac.uk/repository/71602025-05-28T19:46:52.074654Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiafalse |
| dc.title.none.fl_str_mv |
Performance evaluation of nine reference centers and comparison of DNA extraction protocols for effective surveillance of Leishmania-infected Phlebotomine sand flies Basis for technical recommendations |
| title |
Performance evaluation of nine reference centers and comparison of DNA extraction protocols for effective surveillance of Leishmania-infected Phlebotomine sand flies |
| spellingShingle |
Performance evaluation of nine reference centers and comparison of DNA extraction protocols for effective surveillance of Leishmania-infected Phlebotomine sand flies Prudhomme, Jorian Public Health, Environmental and Occupational Health Infectious Diseases SDG 3 - Good Health and Well-being |
| title_short |
Performance evaluation of nine reference centers and comparison of DNA extraction protocols for effective surveillance of Leishmania-infected Phlebotomine sand flies |
| title_full |
Performance evaluation of nine reference centers and comparison of DNA extraction protocols for effective surveillance of Leishmania-infected Phlebotomine sand flies |
| title_fullStr |
Performance evaluation of nine reference centers and comparison of DNA extraction protocols for effective surveillance of Leishmania-infected Phlebotomine sand flies |
| title_full_unstemmed |
Performance evaluation of nine reference centers and comparison of DNA extraction protocols for effective surveillance of Leishmania-infected Phlebotomine sand flies |
| title_sort |
Performance evaluation of nine reference centers and comparison of DNA extraction protocols for effective surveillance of Leishmania-infected Phlebotomine sand flies |
| author |
Prudhomme, Jorian |
| author_facet |
Prudhomme, Jorian Delabarre, Aymeric Alten, Bulent Berberoglu, Umut Berriatua, Eduardo Bongiorno, Gioia Cristovão, José Manuel Davidovich-Cohen, Maya Di Muccio, Trentina Kasap, Ozge Erisoz Fiorentino, Eleonora Kirstein, Oscar D. Kniha, Edwin Maia, Carla Mungan, Mesut Muñoz-Hernández, Clara Nalcaci, Muhammed Kaskan, Gizem Oguz Ozbel, Yusuf Toz, Seray Ozensoy Parreira, Ricardo Platzgummer, Katharina Polat, Ceylan Risueño, José Studentsky, Liora Varol, Gamze Walochnik, Julia Yetişmiş, Kardelen Robert-Gangneux, Florence |
| author_role |
author |
| author2 |
Delabarre, Aymeric Alten, Bulent Berberoglu, Umut Berriatua, Eduardo Bongiorno, Gioia Cristovão, José Manuel Davidovich-Cohen, Maya Di Muccio, Trentina Kasap, Ozge Erisoz Fiorentino, Eleonora Kirstein, Oscar D. Kniha, Edwin Maia, Carla Mungan, Mesut Muñoz-Hernández, Clara Nalcaci, Muhammed Kaskan, Gizem Oguz Ozbel, Yusuf Toz, Seray Ozensoy Parreira, Ricardo Platzgummer, Katharina Polat, Ceylan Risueño, José Studentsky, Liora Varol, Gamze Walochnik, Julia Yetişmiş, Kardelen Robert-Gangneux, Florence |
| author2_role |
author author author author author author author author author author author author author author author author author author author author author author author author author author author author |
| dc.contributor.none.fl_str_mv |
Instituto de Higiene e Medicina Tropical (IHMT) Global Health and Tropical Medicine (GHTM) Laboratório Associado de Translacção e Inovação para a Saúde Global - LA Real (Pólo IHMT) Vector borne diseases and pathogens (VBD) RUN |
| dc.contributor.author.fl_str_mv |
Prudhomme, Jorian Delabarre, Aymeric Alten, Bulent Berberoglu, Umut Berriatua, Eduardo Bongiorno, Gioia Cristovão, José Manuel Davidovich-Cohen, Maya Di Muccio, Trentina Kasap, Ozge Erisoz Fiorentino, Eleonora Kirstein, Oscar D. Kniha, Edwin Maia, Carla Mungan, Mesut Muñoz-Hernández, Clara Nalcaci, Muhammed Kaskan, Gizem Oguz Ozbel, Yusuf Toz, Seray Ozensoy Parreira, Ricardo Platzgummer, Katharina Polat, Ceylan Risueño, José Studentsky, Liora Varol, Gamze Walochnik, Julia Yetişmiş, Kardelen Robert-Gangneux, Florence |
| dc.subject.por.fl_str_mv |
Public Health, Environmental and Occupational Health Infectious Diseases SDG 3 - Good Health and Well-being |
| topic |
Public Health, Environmental and Occupational Health Infectious Diseases SDG 3 - Good Health and Well-being |
| description |
Publisher Copyright: © 2024, Author. All rights reserved. |
| publishDate |
2024 |
| dc.date.none.fl_str_mv |
2024-12 2024-12-01T00:00:00Z 2025-02-05T21:21:10Z |
| dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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publishedVersion |
| dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10362/178507 |
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http://hdl.handle.net/10362/178507 |
| dc.language.iso.fl_str_mv |
eng |
| language |
eng |
| dc.relation.none.fl_str_mv |
1935-2727 PURE: 108808671 https://doi.org/10.1371/journal.pntd.0012543 |
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info:eu-repo/semantics/openAccess |
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openAccess |
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15 application/pdf |
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reponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia instacron:RCAAP |
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FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia |
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RCAAP |
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RCAAP |
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Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) |
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Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) |
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Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia |
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info@rcaap.pt |
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